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目的建立定量评价副溶血弧菌(Vibrio parahaemolyticus,VP)溶血活性的实验方法。方法以大耳白兔红细胞为靶细胞,野生型VP标准株RIMD2210633(J5421)、opaR敲除株、toxR敲除株的菌体细胞为效应细胞,通过调节感染菌量及孵育时间来优化条件,从而建立溶血活性评价模型。结果与结论 VP最佳溶血活性条件为红细胞终浓度5%条件下,用约3×106CFU的菌量感染靶细胞,37℃孵育2.5 h。opaR负调控VP的溶血活性,而ToxR正调控VP的溶血活性。本研究成功建立了定量测定VP溶血活性的方法。
Objective To establish a quantitative method for evaluating the hemolysis activity of Vibrio parahaemolyticus (VP). Methods Erythrocytes were used as target cells. The wild type VP standard strain RIMD2210633 (J5421), opaR knockout strain and toxR knockout strain were used as effector cells. The optimal conditions were optimized by adjusting the bacterial count and incubation time. Thus establishing hemolytic activity evaluation model. RESULTS AND CONCLUSION: The best hemolytic activity of VP was that the target cells were infected with about 3 × 106CFU of bacteria at a final erythrocyte concentration of 5% and incubated at 37 ℃ for 2.5 h. OppaR negatively regulates VP hemolytic activity, whereas ToxR regulates VP hemolytic activity. This study successfully established a quantitative determination of hemolytic activity of VP method.